Method for preparing cosmetic composition with heat-aging inhibitory effect using summer savory extract

Abstract

The present invention relates to a cosmetic composition, and more specifically, to a cosmetic composition having a heat-aging inhibitory effect comprising a summer savory (Satureja hortensis) extract as an active ingredient, and a method for manufacturing the same.

Description

Technology Field

[0001] The present invention relates to a cosmetic composition, and more specifically, to a cosmetic composition having a heat-aging inhibitory effect comprising a summer savory (Satureja hortensis) extract as an active ingredient, and a method for manufacturing the same. Background Technology

[0002] Skin aging is broadly classified into intrinsic aging and extrinsic aging. Intrinsic aging is a biological decline mechanism in which the physiological functions of the skin gradually deteriorate with age, clinically characterized by a decrease in elasticity, roughening of skin texture, and the formation of wrinkles and pigmentation. Extrinsic aging refers to aging phenomena caused by external factors such as heat, light, free radicals, and stress.

[0003] In particular, thermal aging caused by near-infrared radiation and thermal stimulation has been reported to raise epidermal and dermal temperatures, inducing the generation of reactive oxygen species and inflammatory mediating signals, and increasing the expression of collagen-degrading enzymes (such as MMP-1) in the dermis, thereby accelerating the loss of elasticity and wrinkle formation.

[0004] This process involves the activation of TRPV1 ion channels, which are widely present in skin tissues such as keratinocytes, and TRPV1-Ca by thermal stimulation 2+ A mechanism has been proposed in which the protein kinase pathway is initiated and MMP-1 transcription is promoted upon influx.

[0005] Furthermore, thermal stimulation induces the expression of Heat Shock Proteins (HSPs) as part of the intracellular defense system. Since HSPs are designed to repair cellular damage caused by heat stress, increased HSP expression is linked to changes in cellular protein homeostasis resulting from heat stress. This is a significant phenomenon directly associated with structural and functional damage to the skin and can be considered a key indicator in thermal aging research.

[0006] Furthermore, thermal aging is linked to the accumulation of Advanced Glycation End Products (AGEs), leading to structural hardening of the skin. There is accumulating evidence that when non-enzymatic glycation is promoted in high-temperature oxidative environments, matrix proteins such as collagen and elastin become cross-linked and rigid, while receptor-dependent inflammation and oxidative stress are heightened, exacerbating signs of aging.

[0007] Nevertheless, as a significant portion of commercially available cosmetics remain focused on UV protection, research on formulations and manufacturing processes targeting specific pathways of thermal aging, such as the expression of collagen-degrading enzymes in response to heat and infrared radiation, has been relatively limited.

[0008] Meanwhile, summer savory (Satureja hortensis) is a plant of the Lamiaceae family that has traditionally been used as a spice and medicine and is rich in polyphenols and essential oils, including rosmarinic acid, carvacrol, and thymol. Numerous studies have reported that these components exhibit powerful antioxidant, anti-inflammatory, and microbial inhibitory activities, making them noteworthy as bioactive candidates that can contribute to skin barrier protection.

[0009] However, an examination of current research and trends in commercially available cosmetic ingredients reveals that summer savory has been utilized only in a limited capacity, primarily as an antimicrobial agent, preservative, or antioxidant auxiliary material. Attempts to formulate compositions or verify their efficacy by focusing on the specific mechanism of inhibiting skin thermal aging have been insufficient; in particular, no cases of applying summer savory extract to target thermal aging-specific biomarkers have been confirmed in either academic or industrial fields.

[0010] Therefore, despite the abundant polyphenol and essential oil components of summer savory, no prior art or products have been reported that directly link these to the efficacy of inhibiting thermal aging. Consequently, there is a growing demand for the development and commercialization of a method for manufacturing a cosmetic composition containing summer savory extract as an active ingredient that exhibits an inhibitory effect on thermal aging, as well as for the cosmetic composition manufactured thereby. Prior art literature

[0011] Korean Patent Publication No. 10-2009-0132651 (Published Dec. 29, 2009) "Composition for the prevention and treatment of metabolic diseases containing Spanish oregano extract or summer savory extract as an active ingredient" The problem to be solved

[0012] In accordance with the above requirements, the present invention aims to provide a method for manufacturing a cosmetic composition having a heat-aging inhibitory effect containing a summer savory extract as an active ingredient, and a cosmetic composition manufactured thereby, wherein the heat-aging alleviating activity of the active ingredient is expressed and increased through a low-temperature extraction process applying reduced pressure and ultrasonic conditions, and the stability and reproducibility of the composition are enhanced by filtering and freeze-drying the extract, thereby providing quality consistency that allows for easy industrial application. means of solving the problem

[0013] To achieve the above objectives, the present invention provides a method for preparing a cosmetic composition having a heat-aging inhibitory effect containing a summer savory extract as an active ingredient and a cosmetic composition, characterized by comprising: a pretreatment step (S10) of preparing a primary substance by immersing summer savory in purified water; an extraction step (S20) of preparing an extract by low-temperature extraction while treating the primary substance with reduced pressure and ultrasound; and a completion step (S30) of completing a cosmetic composition by filtering, concentrating under reduced pressure, and freeze-drying the extract.

[0014] In addition, the above pretreatment step (S10) is characterized by being a step of preparing a primary substance by immersing summer savory in purified water and adding dry ice.

[0015] Here, the summer savory and purified water are mixed and immersed in a weight ratio of 1:30 to 50, and the dry ice is added in an amount of 5 to 30 weight percent relative to the weight of the purified water.

[0016] And, the extraction step (S20) is characterized by the step of preparing an extract by applying ultrasonic treatment to the primary substance at a reduced pressure of 50 to 150 mbar and at 20 to 60 Khz and 100 to 500 W, and performing low-temperature extraction for 1 to 3 hours. Effects of the invention

[0017] The cosmetic composition according to the manufacturing method of the present invention comprises summer savory extract as an active ingredient and has a heat-aging inhibitory effect, thereby providing an effect of alleviating the decrease in skin elasticity, wrinkle formation, oxidation, and inflammatory reactions caused by heat stimulation. Brief explanation of the drawing

[0018] FIG. 1 is a flowchart schematically illustrating a manufacturing method according to the present invention. Figure 2 is a graph showing the results of the cytotoxicity test. Figure 3 is an image illustrating biomarker expression in HDFn cells. Specific details for implementing the invention

[0019] The following detailed descriptions relating to the present invention refer to the accompanying drawings, which are embodiments in which the present invention may be practiced and are illustrated as examples of such embodiments. These embodiments are described in detail to sufficiently enable those skilled in the art to practice the present invention. It should be understood that various embodiments of the present invention are different but need not be mutually exclusive. For example, specific shapes, structures, and characteristics described herein may be implemented in other embodiments without departing from the spirit and scope of the present invention in relation to one embodiment. Furthermore, it should be understood that the location or arrangement of individual components within each described embodiment may be changed without departing from the spirit and scope of the present invention.

[0020] Accordingly, the following detailed description is not intended to be taken in a limiting sense, and the scope of the invention is limited only by the appended claims, including all equivalents to those claimed therein, provided they are properly described. Similar reference numerals in the drawings refer to the same or similar functions across various aspects.

[0021] The terms used in this invention have been selected based on currently widely used general terms, taking into account their functions within the invention; however, these terms may vary depending on the intent of those skilled in the art, case law, the emergence of new technologies, etc. Additionally, in specific cases, terms have been arbitrarily selected by the applicant, and in such cases, their meanings will be described in detail in the relevant description of the invention. Therefore, the terms used in this invention should be defined not merely by their names, but based on their meanings and the overall content of the invention.

[0022] In the present invention, when a part is described as "comprising" a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but may include additional components.

[0024] The following describes in detail a method for preparing a cosmetic composition having a heat-aging inhibitory effect by including a summer savory extract according to the present invention as an active ingredient.

[0025] At this time, it is clarified that the cosmetic composition of the present invention is manufactured according to the manufacturing method of the present invention and is an invention that shares the same technical concept.

[0027] A method for preparing a cosmetic composition having a heat-aging inhibitory effect by including a summer savory extract of the present invention as an active ingredient comprises a pretreatment step (S10), an extraction step (S20), and a completion step (S30).

[0029] The above pretreatment step (S10) is a step of preparing a primary substance by immersing summer savory in purified water.

[0030] Here, summer savory (Satureja hortensis) is a plant of the genus Satureja belonging to the Lamiaceae family, and is a representative annual herbaceous plant among the various species generally called “savory.”

[0031] In the genus Savory, summer savory and winter savory (Satureja montana) are mainly known. Winter savory is a perennial with woody stems and exhibits a strong aroma and bitter flavor, whereas summer savory is an annual plant with soft, tender stems and a mild flavor, with a balanced distribution of polyphenols and essential oils.

[0032] Although the thermal aging mitigation effect of such summer savory was not previously known, it is possible to manifest and enhance the thermal aging mitigation effect through treatment according to the manufacturing method of the present invention, and this will be explained in detail in the experimental examples to be described later.

[0033] In the present invention, summer savory can be applied in its raw form or in a crushed state, and it is preferable that it be in a well-dried state.

[0034] At this time, the pretreatment step (S10) may be a step of preparing a primary substance by immersing summer savory in purified water and adding dry ice. Here, it is preferable that the purified water be stored at room temperature and be at a temperature of 15 to 25°C.

[0035] Dry ice has the effect of cooling primary substances and sublimates in purified water to generate carbon dioxide bubbles, and the carbon dioxide bubbles replace dissolved oxygen to inhibit oxidative decomposition reactions, thereby reducing the oxidative destruction of components that can provide thermal aging inhibition.

[0036] Furthermore, the localized ultra-low temperature environment created by the contact between dry ice and summer savory effectively crushes the raw material tissue, thereby providing a role that enhances extraction efficiency compared to simply applying low-temperature purified water.

[0037] Furthermore, the above summer savory and purified water are mixed in a weight ratio of 1:30 to 50 and immersed, and the above dry ice can be added in an amount of 5 to 30 weight percent relative to the weight of the purified water.

[0038] Here, when the weight ratio of purified water to summer savory is less than 30, the raw material tissue is not sufficiently swollen, which limits the elution of active ingredients and makes it difficult to uniformly deliver the low-temperature treatment effect by dry ice.

[0039] On the other hand, if the weight ratio of purified water to summer savory exceeds 50, the volume of the immersion solution increases excessively, diluting the cooling and foaming action of dry ice, which may reduce the oxidation inhibition and tissue crushing effects.

[0040] That is, a weight ratio of 1:30 to 50 can be considered a critical condition for stably securing the active ingredients of summer savory and for enhancing oxidation inhibition and tissue breaking effects in the pretreatment step (S10), and it is most desirable for summer savory and purified water to be mixed in a weight ratio of 1:40.

[0041] Meanwhile, if the amount of dry ice added is less than 5% by weight relative to purified water, the dissolved oxygen in the purified water cannot be sufficiently replaced, resulting in a weak inhibitory effect on oxidative decomposition and a localized ultra-low temperature environment not being sufficiently formed, which may limit the breaking of the cell walls of the raw material tissue.

[0042] Conversely, if an excessive amount exceeding 30 weight% is added, heat and mass transfer between the purified water and the summer savory may be inhibited due to rapid bubble generation, or the active ingredients may change unstably due to excessive localized cooling, which may lead to unnecessary energy consumption and reduced efficiency in the process.

[0043] That is, the dry ice input amount of 5 to 30 weight percent is a critical condition that can prevent tissue damage or a decrease in process efficiency while sufficiently securing the oxygen exchange effect and the effect of forming a local ultra-low temperature environment, and it is preferable that the dry ice be added at 20 weight percent relative to the weight of purified water.

[0044] When the pretreatment step (S10) is performed under these desirable conditions, the temperature of the primary material converges to 0 to 5℃.

[0046] The above extraction step (S20) is a step of preparing an extract by treating the primary substance with reduced pressure and ultrasound and extracting it at low temperature.

[0047] At this time, the extraction step (S20) may be a step of preparing an extract by applying ultrasonic treatment to the primary substance at a reduced pressure of 50 to 150 mbar and at 20 to 60 Khz and 100 to 500 W, and performing low-temperature extraction for 1 to 3 hours.

[0048] Preferably, the extraction step (S20) may be a step of preparing an extract by applying ultrasonic treatment to a primary substance at 40 Khz and 300 W at reduced pressure of 100 mbar and low temperature extraction for 2 hours.

[0049] At this time, low-temperature extraction can be achieved by reducing the temperature through the introduction of dry ice as described above, and it is preferable that the temperature be 0 to 5℃.

[0051] The above completion step (S30) is a step of completing the cosmetic composition by filtering, vacuum concentrating, and freeze-drying the extract.

[0052] At this time, filtration is performed to remove impurities or solids, and can be carried out with a filter paper having an average pore size of 1 to 50 µm, and a filter paper having a pore size of 1 to 10 µm is suitable, and Whatman No. 3 (Whatman, USA) can be exemplified therein.

[0053] The above vacuum concentration can be performed with a concentration yield of 40% by weight based on the weight of the raw material extracted, and for example, it may be a process of obtaining 10g of concentrate when extracting 25g of summer savory and then vacuum concentrating.

[0054] The above freeze-drying process can be performed with a cosmetic composition yield of 18.8% by weight based on the weight of the raw extract, i.e., a cosmetic composition yield of 47% by weight based on the concentrate, and, for example, it can be a process of obtaining 4.7g of cosmetic composition for every 10g of concentrate.

[0056] The powdered cosmetic composition produced through the manufacturing method of the present invention can be mixed with various carriers to be formulated as an external skin preparation to provide a thermal aging alleviation effect to the skin, and is not limited thereto.

[0058] Below, the effects of the cosmetic composition prepared by the manufacturing method of the present invention will be examined in detail through examples, comparative examples, and experimental examples.

[0060] Example. Cosmetic composition according to the present invention

[0061] Pretreatment step (S10): 25g of washed and dried raw summer savory was placed in 1L of purified water, 200g of dry ice was added, and the mixture was immersed for 20 minutes while stirring slowly to pretreat and cool the temperature to within the range of 0 to 5℃ to produce a primary substance.

[0062] Extraction step (S20): The primary substance was subjected to reduced pressure at 100 mbar and ultrasonically treated at 40 Khz and 300 W for 2 hours to produce an extract.

[0063] Completion step (S30): The extract was filtered through Whatman No. 3 filter paper, then concentrated under reduced pressure to obtain 10g of concentrate, and the concentrate was freeze-dried to complete a cosmetic composition of 4.7g.

[0065] Comparative Example. Example excluding dry ice treatment

[0066] Pretreatment step (S10): 25g of washed and dried raw summer savory was placed in 1L of purified water and immersed for 20 minutes while stirring slowly to prepare a primary substance. At this time, purified water refrigerated to 4℃ was used to compare temperature conditions equivalent to those in the example.

[0067] Subsequently, the extraction step (S20) and the completion step (S30) were carried out in the same manner as in the example to complete the cosmetic composition.

[0069] The above examples and comparative examples were prepared as liquid samples by mixing purified water and glycerin in a 3:1 volume ratio with a dispersion medium at concentrations of 50, 100, 250, 500, and 1,000 μg / mL.

[0071] Experimental Example 1. Confirmation of non-cytotoxicity

[0072] To confirm cytotoxicity of the samples of the above examples and comparative examples, the effect on cell proliferation was examined.

[0073] 1) Experimental Method

[0074] 1 x 10⁶ HDFn cells in a 24-well plate 5 Cells were cultured at a concentration of cells / mL in a 37℃, 5% CO2 incubator for 24 hours.

[0075] Afterward, the culture medium was removed, and 50 µl of the sample at different concentrations was added to each well, followed by the addition of 450 µl of fresh medium, and then incubated in a 37°C, 5% CO2 incubator for 24 hours.

[0076] Prepare MTT reagent and dilute it to 1 mg / mL. After adding 100 µL to each well, incubate under dark conditions in a 37°C, 5% CO2 incubator for 2 hours. Then, remove the supernatant, add 300 µL of DMSO to each well, and react under dark conditions at room temperature for 5 minutes.

[0077] Finally, 100 µl of the reacted culture solution was added to each well of a 96-well plate, and the absorbance was measured at 540 nm.

[0078] At this time, an untreated sample group (0 μg / mL) was placed as a control group, and the change in absorbance of the sample-treated group relative to the absorbance of the control group was calculated. The absorbance was measured three times for each sample at each concentration and the arithmetic mean was calculated.

[0080] 2) Experimental Results

[0081] The experimental results are as shown in Table 1 below.

[0082] Sample concentration (µg / mL) Example Cell proliferation effect (%) Comparative Example Cell Proliferation Effect (%) 0 100.0 100.0 50 100.5 100.3 100 101.2 100.9 250 101.9 101.3 500 102.1 101.5 1,000 105.5 102.6

[0083] As shown in Table 1 above, the samples of the examples and comparative examples did not exhibit significant cytotoxicity in all concentration ranges, confirming that they can be applied to the skin as cosmetic compositions.

[0085] Experimental Example 2. Confirmation of thermal aging biomarker activity

[0086] Western blot analysis was performed on the samples of the above examples and comparative examples to determine the effect of heat stimulation on the protein stress response.

[0088] 1) Experimental Method

[0089] (1) Cell culture

[0090] HDFn cells were cultured in DMEM medium containing 10% FBS and 1% penicillin-streptomycin. The cultured cells were counted using a hemocytometer and placed in a 6-well plate at a rate of 4 × 10⁶ cells. 5 After dispensing at a concentration of cells / well, the cells were cultured for 24 hours in an incubator at 37°C and 5% CO2.

[0091] Afterward, the culture medium was removed, each well was washed with 1X PBS, serum-free DMEM medium was added, and thermal stress was applied at 44°C for 1 hour. Immediately after the thermal stress, samples at a concentration of 1,000 μg / mL from the examples and comparative examples, as well as the control group (untreated), were added to each well, and the samples were cultured again in a 37°C, 5% CO2 incubator for 24 hours.

[0092] (2) Protein extraction and quantification

[0093] Protein extraction was performed as follows.

[0094] After washing the cultured cells with 1X PBS, 300 µl of RIPA buffer was added to each well and incubated at 4°C for 10 minutes. Cells were scraped off using a cell scraper, and the contents were collected in a 1.5 mL e-tube. Subsequently, the mixture was centrifuged at 12,000 rpm for 10 minutes in a centrifuge at 4°C, and the supernatant was transferred to a new tube.

[0095] Total protein content was quantified using SPECTRA ABS PLUS (Molecular Devices, USA) and adjusted to a concentration of 1,000 μg / mL for Western blot experiments.

[0096] The quantified protein was mixed with 5X protein buffer, heat-denatured at 100°C for 10 minutes, and then slowly cooled over ice to prepare the sample.

[0097] (3) Confirmation of protein marker expression

[0098] Protein analysis was performed using SDS-PAGE with a 10% acrylamide gel. The samples were electrophoresed at 70 V for 20 minutes, and once the stacking gel separated, proteins were separated by electrophoresis at 100 V for 80 minutes. The gels after electrophoresis were transferred using a Trans-Blot instrument at 25 V and 100 mA for 60 minutes.

[0099] Blocking was performed at room temperature for 1 hour using a 5% skim milk solution. Subsequently, the primary antibody was diluted with 5% skim milk at a ratio of 1:1000 and incubated overnight at 4°C.

[0100] After the first reaction, the membrane was washed with 1X TBST buffer, treated with the secondary antibody for 1 hour at room temperature, and finally washed again with 1X TBST and detected the biomarker using ELC (Electrochemiluminescence) solution.

[0101] (4) Biomarker to be detected

[0102] To confirm the inhibitory effect of each sample on thermal aging, MMP 1 and MMP 3 were selected as biomarkers capable of quantifying cellular responses to heat stimulation, and β-actin was selected as a reference protein for balancing and comparing protein quantification between experiments.

[0103] Specifically, the aforementioned Matrix Metalloproteinase (MMP) is an enzyme that degrades collagen and extracellular matrix proteins primarily present in the dermis. It is reported to be a group of proteolytic enzymes that increase particularly sensitively under heat stimulation conditions, and this is utilized as an indicator to quantitatively determine the heat stress response at the cellular level.

[0104] Here, MMP-1 is one of the enzymes whose expression increases first in response to heat stimulation, and it is a primary degrading enzyme that directly breaks down collagen types I and III, which are abundant in the dermis. Therefore, the increase in MMP-1 expression reflects the starting point of skin structural damage caused by heat stress, and whether it is inhibited serves as an indicator that directly proves the effect of alleviating thermal aging.

[0105] MMP-3 is an enzyme that is simultaneously activated with MMP-1 by thermal stimulation; it not only degrades elastin and various extracellular matrix proteins but also acts as a mediator that activates other MMP precursors. In other words, since MMP-3 functions as a marker that triggers a chain reaction of protein degradation following thermal stimulation, a decrease in its expression serves as another direct evidence confirming the inhibition of the progression of thermal aging.

[0106] Meanwhile, heat shock proteins, whose expression is induced by heat stimulation, are expressed to maintain intracellular homeostasis in response to heat stress. It is known that the expression of heat shock proteins is not limited to a simple protective response but is accompanied by negative physiological reactions, such as skin cell death, increased expression of inflammatory mediators, and impaired skin barrier function, in accordance with the objective of maintaining homeostasis.

[0107] These heat shock proteins are known to activate signaling pathways such as MAPK and NF-κB, and since these pathways directly promote the transcription and expression of MMP-1 and MMP-3, consequently, inhibition of MMP-1 and MMP-3 expression can be interpreted as inhibition of heat shock protein expression, thereby strengthening the consistency of the implementation of the heat shock mitigation effect.

[0108] The above-mentioned β-actin is a protein involved in maintaining cell structure and maintains constant expression regardless of heat stimulation, thus serving as a reference for comparing the relative expression levels of other biomarkers.

[0110] 2) Experimental Results

[0111] The experimental results are shown in Figure 3.

[0112] As can be seen from Figure 3, the expression levels of both biomarkers MMP-1 and MMP-3 in the example were significantly reduced.

[0113] In the comparative example, the expression of biomarkers was also reduced compared to the control group, but it showed a significant difference from the example, and since the expression of β-actin was constant in both HaCaT and HDFn cells, it can be confirmed that protein quantification and analysis were performed uniformly between each sample.

[0114] In other words, the composition prepared in the example significantly inhibited the expression of MMP-1 and MMP-3, which are thermal aging indicator proteins that specifically increase due to heat stimulation. This result experimentally demonstrates that the cosmetic composition of the present invention has the effect of directly alleviating the thermal aging response itself caused by heat stress, going beyond simple antioxidant or general cell protective effects.

[0116] Although the present invention has been described together with the accompanying drawings, this is merely one example among various embodiments containing the gist of the present invention, and is intended to enable those skilled in the art to easily implement it. It is clear that the present invention is not limited to the embodiments described above. Accordingly, the scope of protection of the present invention should be interpreted by the following claims, and all technical ideas within an equivalent scope by modification, substitution, replacement, etc., within the scope that does not deviate from the gist of the present invention shall be included in the rights of the present invention. Furthermore, it is clarified that some components in the drawings are provided in an exaggerated or reduced form compared to the actual form to more clearly explain the configuration. Explanation of the symbols

[0117] S10: Preprocessing step S20: Extraction step S30: Completion stage

Claims

Claim 1 A method for preparing a cosmetic composition having a heat-aging inhibitory effect comprising a summer savory extract as an active ingredient, comprising: a pretreatment step (S10) of preparing a primary substance by immersing summer savory in purified water and adding dry ice; an extraction step (S20) of preparing an extract by low-temperature extraction at 0 to 5℃ for 1 to 3 hours while ultrasonically treating the primary substance at 50 to 150 mbar, 20 to 60 Khz, and 100 to 500 W; and a completion step (S30) of completing a cosmetic composition by filtering, vacuum concentrating, and freeze-drying the extract; wherein the cosmetic composition is characterized by inhibiting the expression of MMP-1 and MMP-3, which are heat-aging indicator proteins that increase due to heat stimulation. Claim 2 delete Claim 3 A manufacturing method according to claim 1, characterized in that the summer savory and purified water are mixed and immersed in a weight ratio of 1:30 to 50, and the dry ice is added in an amount of 5 to 30 weight percent relative to the weight of the purified water. Claim 4 delete Claim 5 A cosmetic composition manufactured according to the manufacturing method of claim 1, characterized by having a thermal aging inhibitory effect by inhibiting the expression of MMP-1 and MMP-3, which are thermal aging indicator proteins that increase due to heat stimulation.

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